Method and Formulation for Treating Resistance to Antihypertensives and Related Conditions

ABSTRACT

A method for the prevention or treatment of symptoms of hypertension in a patient who is resistant to antihypertensive effects of an antihypertensive compound administered in the absence of melatonin comprises administering to said patient melatonin in an amount effective to ameliorate or prevent symptoms of hypertension in said patient.

This application is a continuation of U.S. application Ser. No.11/950,039, filed Dec. 4, 2007, which is a divisional of Ser. No.10/169,467 filed Sep. 20, 2002, now U.S. Pat. No. 7,332,177, which was a371 filing of PCT/IL2000/00009, filed Jan. 5, 2000. These priorapplications are incorporated herein by reference.

FIELD AND BACKGROUND OF THE INVENTION

The present invention relates to a method and pharmaceutical formulationfor treating a patient who is resistant to the antihypertensive effectof an antihypertensive compound in absence of melatonin, a method forlowering nocturnal blood pressure in patients who have an abnormalrhythm in blood pressure in the absence or presence of anantihypertensive compound, a method for lowering cortisol levels andprotecting from cardiovascular events, and use of melatonin in themanufacture of medicaments for the stated purposes.

There is a daily variation in blood pressure (circadian blood pressurerhythm) which is characterized by a nocturnal fall and a diurnal rise.The normal pattern of circadian blood pressure rhythm is reversed inelderly people and in those with Cushing's syndrome, those undergoingglucocorticoid treatment, and those with hyperthyroidism, central and/orperipheral autonomic dysfunction (Shy-Drager syndrome, tetraplegia,diabetic or uremic neuropathy etc.), chronic renal failure, renal orcardiac transplantation, congestive heart failure, eclampsia, sleepapnea syndrome, malignant hypertension, systemic atherosclerosis,accelerated hypertensive organ damage (Imai, Abe et al. Journal ofhypertension (supplement) 8:S125-132, 1990) and fatal familial insomnia(Portaluppi, Cortelli et al. Hypertension 23:569-576, 1994). Aless-than-normal decline in nocturnal blood pressure is seen in somehypertensive patients despite treatment with antihypertensive drugs. Aless-than-normal decline in nocturnal blood pressure has been associatedwith excessive cardiovascular complications in hypertensive patients.Patients with impaired nocturnal blood pressure reduction (nondippers)are at increased risk of developing target organ damage (1-4) andnondipper women have been shown to develop more cardiovascular events(5) than their dipper counterparts. The mechanism of the normal fall ofblood pressure during sleep and the pathophysiological mechanismsresponsible for lack of nocturnal fall in blood pressure remain to befully elucidated.

Glucocorticoid hormones play a critical role in a variety of bodilyfunctions. In the basal state, glucocorticoids exert a permissive effecton diverse body functions such as maintenance of blood pressure,euglycemia, and electrolyte and water hemostasis. In humans, cortisol isessential for life. Normally, cortisol secretion from the adrenal glandis rhythmic, with maximal blood levels in the early morning hours, and adecline to half of the peak value in the afternoon. During stress,excretion of cortisol is greatly increased to cope with serious wholebody insult. However, sustained elevation of cortisol in circulation hasdetrimental effects on the immune system and on the ability of the bodyto cope with stress and disease. Most importantly, corticosteroids canprovoke a neurodegenerative process in the hippocampus leading toimpaired memory and cognitive functions. Prolonged exposure of the brainto corticosteroids makes it more vulnerable to degeneration induced byischemia and epilepsy (McEwen, Annals of the New York Academy ofScience, 1994, 746: 145-154). With aging, the basal secretion ofcortisol increases by unknown mechanisms and its peak occurs earlier inthe morning than in young adults (Moreley and Korenman, eds., BlackwellScientific Publications, 1992, pp 70-91). In addition, nocturnalcortisol levels have been found to be higher in coronary patients thanaged-matched healthy subjects (Bragger and Herold, Biological RhythmResearch, 1995, 26: 373). There is an association between hypertensionand high urine cortisol values (Lichtenfeld, Hunt et al, Hypertension,31:569-74, 1998), oral cortisol increases blood pressure in a dosedependent manner (Kelly, Mangos et al, Clin Exp Pharmacol Physiol Suppl25:S51-6, 1998). It has not been previously suggested that there is anassociation between the high cortisol levels and the absence ofnocturnal dip in blood pressure.

Melatonin, the hormone secreted at night from the pineal gland, reachesits peak levels before the onset of the cortisol peak in humans. Theproduction of melatonin declines with age. Also, nocturnal melatoninlevels are lower in coronary patients than in healthy aged-matchedindividuals. However, it has not been suggested that melatonin affectscortisol secretion under normal conditions.

Cardiovascular Effects of Conventional Release Melatonin

Melatonin, the hormone of the pineal gland, is normally secreted atnight and plays a role in the biologic regulation of circadian rhythms,including sleep (Brzezinski, N Engl J Med 1997; 336: 186-195, Penev andZee, Ann Neurol 1997; 42: 545-553). Vasorelaxing action of melatonin (athigh concentrations 10-1000.mu.M) has been observed in rabbit aorta invitro (Satake et al., Gen. Pharmacol., 1991, 22:219-221, and22:1127-1133).

Rodent studies indicate the presence of melatonin receptors in somearterial vessels and it's ability to modulate rat vascular smooth muscletone (Capsoni et al, Neuroreport 1995; 6: 1346-1348, Mahle et al, J BiolRhythms 1997; 12: 690-696). This modulation may be manifested asvasodilatation or vasoconstriction depending on the animal species.

The effects of melatonin on blood pressure and on the humancardiovascular system is complex (Lusardi et al, Blood Press Monit 1997;2: 99-103, Cagnacci et al, 1998; 274: 335-338, Arangino et al, Am JCardiol 1999; 83: 1417-1419, Terzolo at al. J. Pineal Research, 1990, 9:113-124). Bedtime melatonin ingestion (5 mg) for 4 weeks to youngnormotensive subjects caused a decrease in systolic blood pressurethroughout the 24 h period, a decrease in diastolic blood pressurelimited to the second half of the night, a slight lowering of the heartrate during the diurnal hours, and an acceleration during the secondhalf of the night (Lusardi et al, Blood Press Monit 1997; 2: 99-103).The daytime administration of melatonin (1 mg) to young women or menreduced the systolic and diastolic blood pressure within 90 min afteradministration Cagnacci et al, 1998; 274: 335-338; Arangino et al, Am JCardiol 1999; 83: 1417-1419)). The administration of melatonin at 08:00to aged postmenopausal women surprisingly increases their cortisollevels (Cagnacci, Soldani and Yen, L Pineal Res, 22:81-5, 1997).

The effects of long-term (2 months), low dose (2 mg/os daily), timespecified (18:00 h) melatonin administration on endocrine andcardiovascular variables in adult men have also been studied by Terzoloat al. (J. Pineal Research, 1990, 9: 113-124). After treatment, a markedelevation of mean serum melatonin levels were recorded, with asignificant advance of its circadian rhythm. The 24 h patterns ofcortisol and testosterone displayed an anticipation of the morningacrophases at about 1.5 h (not significant) for cortisol and 3 h(significant) for testosterone. Prolactin pattern was unchanged as wellas serum levels of triiodothyronine and thyroxine. Likewise, theresponse of luteinizing hormone (LH), follicle stimulating hormone(FSH), prolactin, thyroid stimulation hormone (TSH) cortisol,adrenocorticotrophin (ACTH) and aldosterone to a stimulation test withgonadotropin releasing hormone (GNRH) thyrotropin releasing hormone(TRH), adrenocorticotrophin (ACTH) and testosterone to human chorionicgonadotrophin (HCG) were also unaffected. The circadian organization ofthe cardiovascular variables, i.e. systolic and diastolic bloodpressure, heart rate, did not show any changes after melatonintreatment.

It is an object of the present invention to the lower cortisol level inhumans and particularly to defer the peak of cortisol in the humancortisol profile. It is a further object of the invention to lower theblood pressure of a patient who is resistant to the antihypertensiveeffect of an antihypertensive compound in absence of melatonin, andespecially to lower the nocturnal blood pressure in non-dippers. It isbelieved that these objects will potentially contribute to decrease inblood pressure, prevention of ischemic attacks and provide prophylacticprotection against the detrimental effects of ischemia on the heart.Other objects of the invention will be apparent from the descriptionwhich follows.

In U.S. Pat. No. 5,700,828, there is described a method for treating orminimizing anoxic or ischemic brain injuries, by administering melatoninto a mammal suffering from an anoxic or ischemic insult, this beingdefined as a trauma that causes a lack of blood flow to the brain and/ora lack of oxygen to the brain. This patent does not suggest thatmelatonin might prevent or ameliorate the anoxic or ischemic insult, perse.

In U.S. Pat. No. 5,849,338, filed Aug. 26, 1997, there is described aunit dosage form for treating vasoconstriction and physiologicalconditions giving rise to it, comprising, in brief, Mg, vitamins C andE, folic acid, Se and melatonin. Melatonin is included only because ofcertain of its properties which were known at the filing date and whichare described in this patent. The entire contents of these U.S. patentsare incorporated by reference herein.

SUMMARY OF THE INVENTION

The above objects may be achieved by the present invention, which in oneaspect, provides a pharmaceutical formulation which comprises, inaddition to at least one carrier, diluent or adjuvant:

melatonin in an amount effective to ameliorate or prevent symptoms ofhypertension developing in a patient who is resistant to theantihypertensive effect of an antihypertensive compound administered inabsence of melatonin; and at least one antihypertensive compound in anamount effective to exert an antihypertensive effect in presence ofmelatonin, in a patient requiring such treatment.

The present invention also provides use of melatonin in the manufactureof a medicament for the prevention or treatment of symptoms ofhypertension in a patient who is resistant to the antihypertensiveeffect of an antihypertensive compound administered in absence ofmelatonin, as well as a method for the prevention or treatment ofsymptoms of hypertension in a patient who is resistant to theantihypertensive effect of an antihypertensive compound administered inabsence of melatonin, which comprises administering melatonin to suchpatient, in an amount effective to ameliorate or prevent symptoms ofhypertension developing in the patient.

According to another aspect, the invention provides use of melatonin inthe manufacture of a medicament for imparting in a patient at least oneeffect selected from improvement in mood and daytime vigilance,postponement of the peak level of cortisol in the patient and potentialprophylactic protection against the detrimental effects of ischemia onthe heart, the medicament being a pharmaceutical formulation whichcomprises melatonin in an amount effective to impart at least one of theabove-stated effects.

According to still another aspect, the invention provides a method forimparting in a patient at least one effect selected from improvement inmood and daytime vigilance, postponement of the peak level of cortisolin the patient and potential prophylactic protection against thedetrimental effects of ischemia on the heart, which comprisesadministering to the patient melatonin in an amount and in a mannereffective to achieve said at least one effect.

The expression “improvement in mood” in the present context is intendedto connote avoidance of mood depression which may be associated withadministration of melatonin in conventional form, i.e. not in controlledrelease form.

Surprisingly, administration of melatonin to humans appears to lowerexcretions rates and diurnal variations. Also, there is a difference inthis respect between controlled- and regular-release melatonin in thatthe controlled release form is able to change and delay the diurnalprofile of cortisol whereas the regular form just suppresses but doesnot shift significantly the time of the peak.

DETAILED DESCRIPTION OF THE INVENTION

The medicament/pharmaceutical formulation may be administered in anyconvenient form, such as one adapted for oral, rectal, parenteral ortransdermal administration. It may be e.g. in unit dosage form. In aparticular embodiment, the melatonin is in the form of a controlledrelease formulation, wherein the melatonin is preferably released at apredetermined controlled rate.

The at least one carrier, diluent or adjuvant may, for example, includeat least one acrylic resin.

The amount of melatonin presently contemplated for use in preventing ortreating hypertension will be the amount found to be effective for thispurpose, presently believed to be, in the case of oral administration,more than 0.5 mg and no more than 100 mg daily, e.g. 0.5-50 mg,preferably 2.5-20 mg, and for parenteral or transdermal administration,between 0.1 and 50 mg. In accordance with the invention, an effectiveamount of melatonin may be formulated e.g. together with an effectivedosage of a antihypertensive drug. The present medicament/pharmaceuticalformulation may comprise also at least one melatonin receptor modifierand/or melatonin profile modifier.

Once the concept of the present invention for treatment or prevention ofhypertension using melatonin is known according to the presentinvention, no inventive skill would be required to ascertain the rangeof effective amounts of melatonin for the present purpose, for variousroutes of administration. Where the pharmaceutical formulation includesat least one antihypertensive agent, this may for example be selectedfrom Diltiazem, Captopril, Atenolol, Benazepril, Enalapril, Valsartan,Metoprolol, Terazosin, Prazosin, Minoxidil, Clonidine, Ramipril andpharmaceutically acceptable salts thereof. The daily dosage rates fororal administration of the exemplified hypertensive compounds, is shownin the following table:

TABLE 1 Antihypertensive Compounds Daily Dosage (mg) Compound Possibleusual Diltiazem HCl 180-300 240 Captopril 12.5-50   12.5 Atenolol 100100 Benazepril HC1  5-20 10 Enalapril Maleate  5-20 10 Valsartan  80-16080 Metoprolol tartarate  95-200 100 Terazosin HCl  1-10 1 Prazosin HC14-64 0.5-5   0.5-1 Minoxidil 5 5 Clonidine HCl 0.15 0.15 Ramipril1.25-5   2.5

The invention will be illustrated by the following Examples.

Example 1

The following ingredients are mixed together and the mixture wascompressed in a 7 mm cylindrical punch, at 2.5 tons, in order to makecontrolled release tablets: Captopril (12.5 mg/tablet), melatonin (5mg/tablet), and Eudragit™ RS100 acrylic resin carrier (Rohm Pharma) andlactose in an approximately 1:1 ratio by weight. While this formulationshould be administered in accordance with a physicians instructions, itis presently contemplated that two such tablets taken two hours beforebedtime would be appropriate.

Example 2

The following ingredients are mixed together and the mixture wascompressed in a 7 mm cylindrical punch, at 2.5 tons, in order to makecontrolled release tablets: Diltiazem (180 mg/tablet), melatonin (5mg/tablet), and Eudragit™ RSPO acrylic resin carrier (Rohm Pharma),lactose and calcium hydrogen phosphate in an approximately 2:1:25 ratioby weight. While this formulation should be administered in accordancewith a physicians instructions, it is presently contemplated that twosuch tablets taken two hours before bedtime would be appropriate.

Experiment 1

The effect of melatonin on blood pressure was determined on a trialpopulation of 52 hypertensive and 130 normotensive elderly patients. Allpatients, who had been insomniacs, were diagnosed according to DSM IV.They consisted of 86 men and 96 women, age 72.+−.9 years. In arandomized, double blind, subjects were given daily either 1, 2 or 5 mgmelatonin in a controlled-release formulation (Circadin™, NeurimPharmaceuticals, Israel), two hours before bedtime, or a placebo ofidentical appearance, for a period of 3 weeks. During the last week ofthe treatment period, BP was assessed at the morning and comparisonswere made between placebo or melatonin treatments, and baseline. Theresults are shown in tables 2 and 3.

TABLE 2 results of Experiment 1 Systolic Systolic Diastolic Diastolicbaseline Treatment baseline Treatment Dose Average SD Average SD P valueAverage SD Average SD Pvalue Hypertensive Patents (>140 mm Hg SystolicBP at baseline) 0 149 5 146 11 0.24 83 6 85 6 0.62 1 145 7 137 9 0.05 824 79 3 0.09 2 147 8 132 9 0.000009 81 6 76 6 0.0064 5 144 5 137 11 0.0482 7 81 6 0.97 Normotensive patients (<140 mm Hg Systolic BP atbaseline) 0 120 11 123 13 0.14 74 7 75 6 0.42 1 121 10 126 16 0.11 75 775 9 0.71 2 122 13 124 15 0.69 75 7 74 8 0.59 5 121 12 124 14 0.16 75 876 9 0.55

Conclusions

Exogenous melatonin administration in the evening decreased daytimesystolic and diastolic in hypertensive elderly subjects. Surprisingly,the administration of the controlled release formulation (1-5 mg) had nosignificant effect in normotensive subjects. It may be noted thatantihypertensive drugs cause a decrease in blood pressure whenadministered to normotensive subjects and that administration of aregular release formulation of melatonin (5 mg) in the evening has beenshown to lower blood pressure in young normotensive subjects throughoutthe 24 h period. (Lusardi et al, Blood Press Monit 1997; 2: 99-103).

Experiment 2

Sixteen elderly patients with essential hypertension were studied.Twenty-four hour ambulatory blood pressures were measured in allpatients. Patients were defined as dippers (n=8) or nondippers (n=8)according to nocturnal fall in mean arterial pressure. 24-hours urinewas collected in two collections, one during daytime, and one duringnighttime. Urinary excretion of the main melatonin metabolite6-sulfatoxymelatonin (6SMT) was determined by ELISA assay in duplicates.Both groups were similar in regard to age and sex. Mean arterialpressure decreased by 10.2% during nighttime in the dippers andincreased by 8% in the nondipper patients Urinary 6SMT increased by 240%during sleep, from 3.28.+−.0.87 (units) during daytime to 8.19.+−.1.68(units) during nighttime (p<0.05) in the dippers, whereas it remainedunchanged in the nondippers (2.31.+−.0.68 (units) during daytime and2.56.+−.0.79 (units) during nighttime). Results are shown in table 3.

TABLE 3 results of Experiment 2 Dippers Nondippers (n = 8) (n = 8) Day3.28 ± 0.87 2.31 ± 0.68 Night 8.19 ± 1.68 2.56 ± 0.79

Conclusions

Nondipper hypertensive patients exhibit blunted nocturnal melatoninsecretion. Thus, exogenous melatonin may play a role in the circadianrhythm of blood pressure.

Investigation of the Effect of Melatonin on Cortisol Profile and Mood

The following experiments were performed in a double-blind, placebocontrolled crossover fashion. Each patient received all three kinds oftablets (placebo, regular release and controlled release) but in randomorder not known to him or the staff.

Experiment 3

Administration of melatonin (2 mg) in a controlled release formulation(SR-Mf), once daily at 10 PM, for one week, to eight healthy elderlypersons suffering from insomnia, resulted in a significant increase intheir sleep efficiency but not sleep latency. (Sleep efficiency is theamount of time spent asleep from total time in bed; sleep latency is thetime taken to fall asleep from first lights-off). On the other handtreatment of the same individuals with melatonin (2 mg) in a regularrelease formulation (RM) did not improve sleep efficiency but shortenedsleep latency compared to placebo treatment of the same subjects. Theseresults can be explained by the short half-life of melatonin in theblood. Namely, the controlled release formulation produces lower bloodlevels of the hormone for extended periods of time and thus its effectsmay start slowly but may be significant later on during the night.

The cortisol level in these patients was assessed by the urinaryexcretion of the hormone at 2 hours intervals over a 24 hour period. Inthe placebo treatment group, patients displayed a cortisol rhythm whichreached its peak at 8:36 AM and the cortisol then declined as is knownfor subjects above 40 years of age (see Sherman et al., Journal ofClinical Endocrinology and Metabolism 1985, 61: 439). The mean 24 hourexcretion rate/hour (which approximated blood concentrations) of thecortisol in urine in the control group was 3.2 microgram/hour. Theamplitude of the rhythm (i.e. maximal deviation of the mean 24 h tomaximum or minimum excretion rate) was 1.8 μg/hour.

After treatment for 1 week with the regular release melatonin theoverall amount of cortisol excreted was reduced. The mean 24 hourexcretion rate decreased to 2.5 μg/hour and the amplitude decreased to1.0 μg/hour. In addition there was a slight backwards shift in the timeof the peak, which occurred at 8:27 AM. Anticipation of the cortisolrhythm after administration of regular release melatonin is compatiblewith observations made by Terzolo at al., J. Pineal Research, 1990, 9:113-124. However, decrease in mean 24 hour levels and amplitude of thecortisol rhythm was not observed by Terzolo.

After one week's treatment with controlled release melatonin, it wasfound that like the regular melatonin, secretion of cortisol wasattenuated (mean 24 h rate was 2.5 μg/hour) and the amplitude 1.2μg/hour as with the regular release), but the peak was delayedsignificantly to later in the day and occurred at 12:06 PM. Thus, thepeak was delayed by administration of controlled release melatonininstead of being the same or slightly advanced. The same cortisolprofile was also found in these patients after 1 month's treatment withthe controlled release formulation (mean 24 hour excretion 2.5 μg/hour,amplitude 1.0 μg/hour and peak time 12:08 hours).

Conclusions

These results show that the response of the body to melatonin is notobvious: the body reads the melatonin profile and not just the fact thatit is present at some time. Interestingly, in humans younger than 40years, the cortisol rhythm is also delayed compared to older individuals(Sherman et al., loc cit). Hence, the cortisol profile generated in theelderly after the controlled release melatonin treatment is similar tothat in younger individuals.

Discussion

It has recently been found that in coronary patients, melatonin at nightis low whereas cortisol levels are high (Bragger and Herold, BiologicalRhythm Research, 1995, 26: 373). It should be noted that cortisol is astress hormone, and its high levels in the morning may be linked to theincreased prevalence of heart attacks in the morning hours. The presentexperiment shows that administration of regular release melatonin canlower cortisol production but that administration of controlled releasemelatonin both lowers the cortisol level and delays its peak and thuscan potentially lower the risk for an ischemic attack during the morninghours.

Experiment 4

This experiment was performed on 10 young healthy males age 26-30. Theyreceived one controlled-release (SR-MI) or regular release (RM) tabletcontaining melatonin (2 mg) or placebo per day with one day washoutbetween treatments. The tablets were taken at 11:00 AM and the subjectswere asked to sleep between 12:00-15:00 hours (i.e., between 12:00-3:00PM). Mood was assessed by Lader-Bond visual analog scale questionnairesbefore and after the sleep. The results indicated that regular melatonin(2 mg) significantly shortened nap sleep latency and increased sleepefficiency. The controlled release formulation also had similar effects.However, the regular release form produced feelings of hostility andsleepiness whereas the controlled release form had no negative effect onmood. These data also indicate that the effects of melatonin on mooddepend on the profile generated. It should be noted that the lack ofeffect on mood cannot be explained by the lower concentrations ofmelatonin generated in he blood by the controlled release formulationsince similar concentrations of melatonin (regular) have been shown byseveral studies to affect mood and sleepiness. Hence, both the timingand pattern of melatonin administration are important in affectingphysiological parameters. The same dose given at different times or indifferent patterns may have different effects.

While particular embodiments of the invention have been particularlydescribed hereinabove, it will be appreciated that the present inventionis not limited thereto, since as will be readily apparent to skilledpersons, many variations and modifications can be made. Such variationsand modifications which have not been detailed herein are deemed to beobvious equivalents of the present invention. For example, structuralanalogs of melatonin which substantially imitate the function ofmelatonin in the human body are deemed to be obvious chemicalequivalents of melatonin. The essential concept, spirit and scope of thepresent invention will be better understood in the light of the claimswhich follow.

1-10. (canceled)
 11. A method for imparting in a patient improvement inmood and daytime vigilance which comprises administering to a patient inneed of such improvement a controlled release pharmaceutical formulationadapted for oral, parenteral or rectal administration comprising aneffective amount of melatonin.
 12. The method of claim 11, wherein saidformulation releases melatonin over a predetermined time period.
 13. Themethod of claim 11, wherein said formulation releases melatoninaccording to a profile which simulates the nocturnal profile in plasmaof a human having a normal endogenous melatonin nocturnal profile.14-19. (canceled)
 20. The method of claim 11, wherein said formulationis in unit dosage form containing an amount within the range of 0.1milligrams to 50 milligrams melatonin per dosage unit.
 21. The method ofclaim 11, wherein said formulation is in particulate form comprisingcoated particles, and the desired controlled release properties areachieved by at least one of the following features: (a) variation in theparticle size of the melatonin; (b) use of at least two coatingmaterials which dissolve at different rates in the human body; or (c)variation in the thickness of at least one coating material, wherebyparticulate melatonin is coated with different thicknesses of saidcoating material(s) which dissolve at different rates in the human body.22. The method of claim 11, wherein said formulation comprisesparticulate melatonin coated with at least one polymeric coatingmaterial.
 23. The method of claim 21, wherein said formulation comprisesparticulate melatonin coated with at least one polymeric coatingmaterial.
 24. The method of claim 11, wherein said formulation furthercomprises at least one melatonin receptor modifier or melatonin profilemodifier.
 25. The method of claim 11, wherein said formulation furthercomprises at least one additional medicament selected frombenzodiazepine melatonin receptor modifiers, benzodiazepine melatoninprofile modifiers, beta-blockers, calcium channel blockers and serotoninuptake inhibitors.
 26. The method of claim 11, wherein said formulationfurther comprises at least one acrylic resin.